ORIGINAL ART ICLE
Evaluation of transgenic event CBH 351 (StarLink) cornin broiler chicks
Chisato YONEMOCHI,1 Hirokazu FUJISAKI,1 Chisato HARADA,1 Toyoko KUSAMA2 and Michito HANAZUMI1
1

Japan Scientific Feeds Association, Chiyoda-ku and 2National Fertilizer and Feed Inspection Station,
Saitama-shi, Japan

ABSTRACTThe influence of transgenic event CBH 351 (StarLink; SL)-derived hybrid corn on the growth performance, health conditionand physiological function in broiler chicks, as well as the possible transfer of the cry9C gene and Cry9C protein to blood,
liver and muscle were examined in comparison with chicks fed on a diet with non-transgenic corn (SL-F). Bodyweight gain
and feed conversion ratio in the chicks fed on a diet with SL were significantly greater than in chicks fed on a diet with
SL-F during the starter phase (0–3 weeks of age), but this significant difference disappeared during the finisher phase(4–7 weeks of age). No abnormalities in health condition in either SL or SL-F groups were observed, and livability did not
differ significantly between SL and SL-F groups. Moreover, no significant differences in serum biochemical and hematological values, histopathological observation and necropsy findings were observed between SL and SL-F groups at the endof the experiment. The cry9C gene and Cry9C protein were not detected in blood, liver and muscle of chicks at 3, 5 or
7 weeks of age. The results indicate that feeding SL does not influence growth performance, health condition or physiological function in broiler chicks, and the cry9C gene and Cry9C protein are not transferred to the blood, liver and musclesof broilers.

KEYWORDS: broiler chicks, growth performance, StarLink corn, transfer of cry9C and Cry9C.

INTRODUCTION

for Safety Assessment of Feed Produced by the
Recombinant DNA Techniques’ (Ministry ofAgriculture, Forestry and Fisheries, MAFF 1996).
These guidelines were prepared on the basis of concepts and principles (namely, substantial equivalence)for safety evaluation of foods derived by modern
biotechnology, which were presented by the OECD
(1993). At the present time, GM crops that have been
confirmed for their safety and approved for the use asa feed ingredient by the Ministry of Agriculture,
Forestry and Fisheries total 30 varieties, including
corn, soybean, rapeseed, cottonseed and beet.
In October 2000, social unease developed when
it was discovered that part of the food and feed

Many varieties of crop plants with improved agronomic characteristics (e.g. insect resistance, herbicide
tolerance, disease resistance and climate tolerance)and enhanced consumer benefits (e.g. better taste andtexture, longer shelf-life, low-allergen potential andmore nutritious) have been developed through agricultural biotechnology. Because genetically modified(GM) crops bring about a profit to the growers
through prevention of crop losses by pest control or areduction in the amount of herbicide applied to thefield, the harvested area of GM crops has increased,
mainly in the USA, Argentina and Canada, every year
since 1996. Genetically modified crops with insect
resistance and herbicide tolerance are used in animal
feeds, however, only GM crops approved by regulatory agencies are commercialized in each country.
In Japan, the safety assessment of GM crops as feed
ingredients is conducted according to the ‘Guidelines

Animal Science Journal (2002) 73, 221–228

Correspondence: Chisato Yonemochi, Japan ScientificFeeds Association, Chiyoda-ku 101-0062, Japan. (Email:yone@kashikyo.or.jp)
Received 31 October 2001; accepted for publication 13
December 2001.

221

C. YONEMOCHI et al.

distributed in Japan contained StarLink (SL). At thattime, SL was not approved by regulatory agencies inJapan for food and feed, but was approved by USA
regulators for feed. StarLink, an insect-resistant cornvariety, contains a novel gene from a naturally occurring soil bacterium, Bacillus thuringiensis subsp. tolworthi, which codes for a form of the Bt toxin known
as Cry9C protein (Schepf et al. 1998). Aventis
CropScience (MN, USA) subjected SL to safety assessments, but a feeding experiment in domestic animals
has not been published.
In this study, the effect of SL on growth performance, health condition and physiological function
was examined in broiler chicks. We also wanted todetermine if the cry9C gene and Cry9C protein contained in SL could be transferred to blood, liver andmuscles of the chicks.

Table 1 Composition of diets

MATERIALS AND METHODSCorn grain
The transgenic corn used was event CBH 351-derived
hybrid (SL) and was purchased from Aventis
CropScience. This corn grain contained 89.3% of thegrain carrying the cry9C gene.
The control non-transgenic corn (SL-F) used was acommercial corn imported from the USA. In the SL-F,
the cry9C gene was not detected by polymerase chain
reaction (PCR), and the Cry9C protein was notdetected by an enzyme-linked immunosorbent assay(ELISA). Before formulating the diet, both SL and
SL-F were ground through 1-mm sieves.

Starter diet†

Finisher diet‡Corn§
Soybean protein
concentrate
Dehulled soybean meal
Fish meal (crude
protein 65%)
Soybean oil
Calcium carbonate
Dicalcium phosphate
Sodium chloride
Vitamin B premix¶
Vitamin ADE premix††
Trace mineral premix‡‡
L -Lysine HCL
DL-Methionine
L-Tryptophan
L-Threonine
L-Arginine
Calculated analysis§§
Crude protein (%)
Metabolizable energy(Mcal/kg diet)
Calcium (%)
Non-phytate
phosphorus (%)
Available lysine (%)
Available
methionine (%)
Available threonine (%)

70.00
7.00

70.00
–

13.27
6.00

20.12
5.00

1.00
0.72
1.30
0.20
0.10
0.10
0.10
0.13
0.05
0.01
0.01
0.01

2.50
0.73
1.08
0.20
0.10
0.10
0.10
0.07
–
–
–
–

20.9
3.15

19.0
3.19

0.97
0.49

0.80
0.44

1.08
0.45

0.92
0.37

0.72

0.65

†Starter diets were fed to chicks from 0 to 3 weeks of age.
‡Finisher diets were fed to chicks from 4 to 7 weeks of age.
§Transgenic corn (StarLink) was used in the diets for theexperimental group and non-transgenic corn was used in the diets
for the control group. ¶Contained (g/kg of premix): thiamine
nitrate 2.0, riboflavin 10.0, pyridoxine hydrochloride 2.0, nicotinic
amide 2.0, D -calcium pantothenic acid 4.35, choline chloride 138.0,
folic acid 1.0 and defatted rice bran. ††Contained (IU or mg/kg
of premix): vitamin A 10 000 IU, vitamin D3 2000 IU, DL-atocopherol acetate 10 mg and defatted rice bran. ‡‡Contained(g/kg of premix): manganese 80, iron 6.0, copper 0.6, zinc 50,
iodine 1.0 and defatted rice bran. §§Based on Standard Table ofFeed Composition in Japan (1995) (Ministry of Agriculture,
Forestry and Fisheries Research Council Secretariat, MAFF 1995).

Diet
The composition of the starter and finisher diets isshown in Table 1. Chicks were fed the starter diet for
3 weeks (0–3 weeks of age; starter phase) followedby the finisher diet for the subsequent 4 weeks(4–7 weeks of age; finisher phase). The diet for the
SL-F group contained 70% SL-F, and the diet for SL
group contained 70% SL. All diets were formulated tomeet or exceed recommended nutrient requirements(Ministry of Agriculture, Forestry and Fisheries
Research Council Secretariat, MAFF 1997) based on
the Standard Table of Feed Composition in Japan(1995) (Ministry of Agriculture, Forestry and Fisheries
Research Council Secretariat, MAFF 1995).

local hatchery and were randomly allotted into eightpens according to sex with 16 chicks each (two treatments, four replications). The diets and tap water
were available ad libitum. In the starter phase, chicks
were maintained in electrically heated batteries in awindowless poultry house. The temperature of theroom was set at 34°C and gradually decreased to 25°C.
To prevent mutual contamination, the battery for the
SL group was separated by a vinyl sheet curtain fromthat for the SL-F group. In the finisher phase, chicks
were maintained in wire-mesh cages in an open-type
poultry house. To prevent mutual contamination, theExperimental design
A total of 256 1-day-old Chunky chicks (equal
number of males and females) were obtained from a

Animal Science Journal (2002) 73, 221–228

Ingredients (%)

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Evaluation of StarLink corn in broilers

SL and SL-F groups were separately housed at either
end of the room.

and SL-F groups. Individual blood, liver and muscle
samples were collected.
For detection of the cry9C gene by PCR, DNA was isolated from the heparinized bloods using a commercial
DNA extraction kit (Dr. GenTLE; Takara Shuzo Co. Ltd,
Kyoto, Japan) according to the manufacturer’s protocol. Liver and muscle were frozen in liquid nitrogen,and finely ground before DNA extraction. DNA was isolated from the ground tissues using a commercial DNA
extraction kit (Isotissue; Nippon Gene Co. Ltd, Toyama,
Japan) according to the manufacturer’s protocol.
The PCR amplification was conducted in a Takara
PCR Thermal Cycler (MP model; Takara Shuzo Co.
Ltd) according to the the conditions recommended by
Aventis CropScience and by Shen et al. (1999) respectively. The amplified fragments were separated on 3%
agarose gel. Following separation of the fragments,
the gels were stained with ethidium bromide solution(0.5 mg/mL). The gel was photographed with a CCD
camera (AE-6915 model; Atto Co. Ltd, Tokyo, Japan)
under UV irradiation. The PCR amplification was
performed using two sets of primers. The PCR primer
set specific for amplification of a 379-bp cry9C generegion in transgenic DNA was designed by Aventis
CropScience. The PCR primer set specific for amplification of a 1075-bp cytochrome b region in mtDNA ofchicks was designed by Shen et al. (1999).
For the detection of the Cry9C protein by ELISA,
blood was mixed with an equal volume of sterilewater. After the sample was homogenized, the sample
was centrifuged at 500 g for 5 min. The supernatant
was used as the analytical material. Liver and muscle
were frozen in liquid nitrogen and finely ground.
After mixing with five parts extraction buffer, thesamples were homogenized. The samples were centrifuged at 500 g for 5 min for muscle, and at 7000 g for
10 min for liver. The supernatants were diluted 4–40fold with water, and the diluted samples were used as
the analytical materials.
The determination of Cry9C protein in the analyticalmaterials was performed using a commercial GMO
Bt9 maize test kit and Cry9C standard solution(Strategic diagnostics Inc., Delaware, USA) according
to the manufacturer’s protocol. After the reaction was
stopped, the extinction was measured at 450 nm witha microplate reader (Spectra classic model; TECAN
Austria GmbH, Salzburg, Austria).

Growth performance
Individual bodyweight was determined at 0, 3, 5 and7 weeks of age. Feed intake and livability were recorded
every day for each replication. The feed conversion
ratio was calculated for the starter phase, the finisher
phase and the entire experimental period. The individual health status of the chicks was monitored every day.

Serum biochemical, hematological andhistopathological examination
At the end of the experiment, 20 chicks comprising anequal numbers of males and females were randomly
chosen from the SL and SL-F groups. The chicks
were anesthetized with ether and then humanely
killed.
For the serum biochemical examination, bloods
were collected from the jugular vein, and sera
were separated. Lactate dehydrogenase (LDH), glutamic-oxaloacetic transaminase (GOT), alkaline phosphatase (ALP), blood urea nitrogen (BUN), neutral
lipid (NL), total cholesterol (T-Chol), total protein (TP),albumin (Alb) and glucose (Glu) were determined byan autoanalyzer (JCB MB8 model; Nihon Denshi Co.,Tokyo, Japan). Globulin (Glob) and the Alb/Glob(A/G) ratio were calculated on the basis of TP and Alb.
For the hematological examination, blood samples
were collected from the jugular vein. Hemoglobin(Hb) (azido-methemoglobin method), hematocrit (Ht)(capillary method), red blood cell count (RBC)(Coulter Counter, model ZF; Coulter Electronics,
Florida, USA), and white blood cell (WBC) (indirect
method) were determined (Anon 1980).
For the histopathological examination, tissue
samples of brain, lung, heart, liver, kidney, spleen,pancreas, duodenum and jejunoileum were collected.
The tissue samples were fixed, thin sections wereprepared, stained with hematoxylin and eosin, andexamined with a light microscope.

Detection of the cry9C gene and Cry9C
protein in blood, liver and muscle
At 3 and 5 weeks of age, a total of 16 chicks comprising an equal number of males and females werechosen from the SL and SL-F groups. In this case, one
chick, with an average bodyweight, was selected fromeach replicate group. At the end of the experiment, a
total of 20 chicks comprising an equal number ofmales and females were randomly chosen from the SL

Animal Science Journal (2002) 73, 221–228

Statistical analysis
Data was analyzed by a two-way ANOVA (Yoshida1985), which included the main effect of diet and sex,223

C. YONEMOCHI et al.

and the interaction between diet and sex. The significance of differences between means was tested at thelevel of P < 0.05. The data for chicks that were killed
for sampling at 3 and 5 weeks of age, or that died orwere culled during the experimental period, were
removed from the statistical analyses of bodyweightgain and feed conversion ratio.

male in the SL-F group died by accident and one
female in the SL group died of ascites. In addition, one
male in the SL group was culled because of perosis.
There were no significant differences in livability
between the SL and SL-F groups.
During the starter phase, bodyweight gain in the SL
group was significantly greater than that in the SL-F
group, and feed intake of the SL group was less thanthat of the SL-F group. Therefore, the feed conversion
ratio of the SL group for the starter phase was significantly greater than that of the SL-F group. However,during the finisher phase and over the entire experimental period, bodyweight gain, feed intake and feed
conversion ratio were not significantly different

RESULTS
Growth performance
Bodyweight gain and livability, and feed intake andfeed conversion ratio are presented in Tables 2 and 3,
respectively. During the experimental period, one

Table 2 Bodyweight gain and livability of chicks during the starter phase (0–3 weeks
of age), finisher phase (4–7 weeks of age) and the entire experimental period

Bodyweight gain (g)DietSL-F group
SL group
Sex
Male
FemalePooled SE
Probabilities
Diet
Sex
Diet ¥ sex

Livability (%)(0–7 weeks)

0–3 weeks

4–7 weeks

0–7 weeks

611
648

1903
1931

2514
2579

99.2
98.5

647
613
30

1953
1881
90

2621
2473
108

98.5
99.2
2.4

< 0.01< 0.01
NS

NS
NS
NS

NS
< 0.01
NS

NS
NS
NS

Each value is the mean of four replications. Transgenic corn (StarLink) was used in the diets
for the experimental group (SL) and non-transgenic corn was used in the diets for the control
group (SL-F). NS, not significant (P > 0.05).

Table 3 Feed intake and feed conversion ratio of chicks during the starter phase (0–3 weeks of age), finisher phase (4–7 weeks
of age) and the entire experimental period

Feed intake (g)Diet
SL-F group
SL group
Sex
Male
FemalePooled SE
Probabilities
Diet
Sex
Diet ¥ sex

Feed conversion ratio

0–3 weeks

4–7 weeks

0–7 weeks

0–3 weeks

4–7 weeks

0–7 weeks

820
796

3781
3872

4601
4668

1.34
1.23

1.99
2.01

1.83
1.81

824
793
26

3914
3739
150

4738
4531
165

1.27
1.30
0.06

2.01
1.99
0.06

1.82
1.82
0.04

0.03
0.01
NS

NS
0.02
NS

NS
0.01
NS

< 0.01
0.01
NS

NS
NS
NS

NS
NS
NS

Each value is the mean for four replications. Transgenic corn (StarLink) was used in the diets for the experimental group (SL) and nontransgenic corn was used in the diets for the control group (SL-F). NS, not significant (P > 0.05).

Animal Science Journal (2002) 73, 221–228

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Hematological values

between the SL and SL-F groups. The interactions
were not significant for livability, bodyweight gain,
feed intake and feed conversion ratio in the starter
phase, the finisher phase or the entire experimental
period.

The hematological values for both groups are presented in Table 5. No significant differences between
SL and SL-F groups were observed for the values of
Hb, Ht, RBC and WBC. However, RBC was significantly higher in females than in males. The interactions were not significant for the hematological values
examined.

Serum biochemical values
The serum biochemical values are presented inTable 4. No significant differences between SL and
SL-F groups were observed for LDH, GOT, ALP, BUN,
NL, T-Chol, Glu, TP, Alb and Glob. However, LDH,
GOT, TP, Alb and Glob were significantly higher infemales than in males. Conversely, T-Chol and Glu
were significantly higher in males than in females. For
other measurements, significant differences by sex
were not observed.

Histopathological observations
Histopathological observations were performed on
samples from males and females of both SL and SL-F
groups. No significant changes in tissue samples ofbrain, lung, heart, liver, kidney, spleen, pancreas, duodenum and jejunoileum of chicks were observed inTable 4 Serum biochemical values for chicks at the end of the experiment

Diet
SL-F group
SL group
Sex
Male
FemalePooled SE
Probabilities
Diet
Sex
Diet ¥ sex

LDH(IU/dL)GOT(IU/dL)ALP(IU/dL)

BUN(mg/dL)NL(mg/dL)

T-Chol
(mg/dL)Glu(mg/dL)TP(g/dL)

Alb(g/dL)

Glob(g/dL)

A/G

1411.01476.6

249.9
255.2

6231.15949.8

0.42
0.60

99.1
91.1

116.5
109.7

233.0
226.9

3.65
3.60

1.39
1.39

2.26
2.21

0.62
0.63

1225.31662.3
402.7

240.2
264.9
31.9

6530.45650.4
805.4

0.44
0.59
0.31

94.3
95.9
22.2

117.9
106.8
10.9

235.1
224.7
14.6

3.51
3.73
0.23

1.34
1.44
0.13

2.17
2.29
0.15

0.62
0.63
0.06

NS
< 0.01
NS

NS
0.01
0.05

NS
NS
NS

NS
NS
NS

NS
NS
NS

NS
< 0.01
NS

NS
0.02
NS

NS
< 0.01
NS

NS
0.01
NS

NS
0.01
0.04

NS
NS
NS

Values are means, n = 10 chicks/sex/group (i.e. total n = 40). Transgenic corn (StarLink) was used in the diets for the experimental group(SL) and non-transgenic corn was used in the diets for the control group (SL-F). A/G, albumin/globulin ratio; Alb, albumin; ALP, alkaline
phosphatase; BUN, blood urea nitrogen; Glob, globulin; Glu, glucose; GOT, glutamic-oxaloacetic transaminase; LDH, lactate dehydrogenase;
NL, neutral lipid; NS, not significant; T-Chol, total cholesterol; TP, total protein.

Table 5 Hematological values for chicks at the end of the experiment

Diet
SL-F group
SL group
Sex
Male
FemalePooled SE
Probabilities
Diet
Sex
Diet ¥ sex

Hb(g/dL)

Ht(%)

RBC(¥ 106/mm3)

WBC(¥ 103/mm3)

8.9
8.5

31.7
31.5

3.34
3.21

30.0
25.8

9.0
8.4
1.1

30.9
32.4
2.7

3.18
3.38
0.25

26.3
29.5
7.6

NS
NS
0.03

NS
NS
NS

NS
0.03
NS

NS
NS
NS

Values are means, n = 10 chicks/sex/group (i.e. total n = 40). Transgenic corn (StarLink) wasused in the diets for the experimental group (SL) and non-transgenic corn was used in thediets for the control group (SL-F). Hb, hemoglobin; Ht, hematocrit; NS, not significant. RBC,
red blood cell count; WBC, white blood cell.

Animal Science Journal (2002) 73, 221–228

225

C. YONEMOCHI et al.

(a)

(b)

Fig. 1 Detection of polymerase chain reaction products specific for (a) the cytochrome b region and (b) cry9C gene in the liverof broiler chickens. Lanes 1–8, livers of broiler chickens from the control group. Lanes 9–16, livers of broiler chickens from theexperimental group. N, no liver DNA; P, positive control.

either group (data not shown). There were no abnormalities observed in the main organs of the chicks.

eventually chicks in the experimental group continued to take a diet containing about 63% SL fromthe start to the end of the experiment.
The present data showed that bodyweight gain andfeed conversion ratio in the SL group during thestarter phase were significantly greater than in the
SL-F group, but this significant difference was notpresent during the finishing phase or over the entire
experimental period. There may be two possible
reasons for this. First, there may be a difference in the
nutritive value between SL and SL-F, influencing thenutritive value of the diets. Second, the battery of
the SL-F group was separated from the battery of theSL group by a vinyl sheet curtain in a windowless
poultry house to prevent mutual contamination, possibly resulting in a difference in the environmental
conditions between the two batteries.
In order to examine the first possibility, the crude
protein content and amino acid composition of eachdiet used in this study were determined by a standard
method. In addition, apparent metabolizable energy
values of each diet were determined by the chromiumoxide-indicator method using three 35-day-old broiler
chicks for each diet. As shown in Table 6, there wereno significant differences in crude protein content,amino acid composition and apparent metabolizableenergy value between SL and SL-F diets for both
starter and finisher diets. Therefore, it was assumed
that the significant differences in bodyweight gain andfeed conversion ratio between SL and SL-F groups
during the starter phase were not caused by SL intake,Possibility of cry9C gene and Cry9C proteintransfer to blood, liver and muscles
At 3, 5 and 7 weeks of age, the cry9C gene and Cry9C
protein were not detectable in blood, liver and muscles in SL and SL-F groups as determined by PCR
and ELISA, respectively (data not shown). In allcases, DNA strands of the 1075-bp cytochrome b
region were amplified by PCR from DNA obtained
from blood, liver and muscles in both groups, whereas
DNA strands of the 379-bp cry9C region were
not amplified. The PCR products amplified from liver
DNA at 3 weeks of age are shown in Figure 1.
The same results were obtained for samples fromblood, liver and muscles at 5 and 7 weeks of age(data not shown). These results indicate that the cry9C
gene and Cry9C protein contained in SL are nottransferred to blood, liver and muscle of chicks afteringestion.

DISCUSSION
The influence of SL on growth and health, as well as
the possibility of transfer of the cry9C gene and Cry9C
protein to blood, liver and muscle were examined.
The diets used in this study contained 70% SL or SL-F,
which is accepted as the upper limit of the formula for
broiler chicks from the viewpoint of animal nutrition.
Since SL contained 89.3% corn grain carrying cry9C,

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Table 6 Crude protein content, apparent metabolizable energy value and animalacid composition of diets

Starter diet

Crude protein (%)
Metabolizable energy (Mcal/kg diet)Arginine (%)
Glycine (%)
Serine (%)
Histidine (%)
Isoleucine (%)
Leucine (%)
Lysine (%)
Methionine (%)
Cystine (%)
Phenylalanine (%)
Threonine (%)
Tryptophan (%)
Valine (%)

Finisher diet

SL-F diet

SL dietSL-F diet

SL diet

20.3
3.09
1.15
0.90
0.92
0.52
0.77
1.72
1.11
0.43
0.30
0.90
0.76
0.23
0.85

20.6
3.11
1.14
0.91
0.93
0.53
0.77
1.65
1.10
0.43
0.31
0.88
0.77
0.24
0.86

18.7
3.12
1.09
0.89
0.90
0.50
0.75
1.64
1.03
0.36
0.28
0.88
0.72
0.21
0.84

18.4
3.11
1.01
0.82
0.83
0.49
0.70
1.52
0.99
0.37
0.29
0.81
0.67
0.20
0.77

Data are measured values. Transgenic corn (StarLink) was used in the experimental diet (SL)and non-transgenic corn was used in the control diet (SL-F).

but, more likely, by environmental factors. This supposition may be supported by the fact that bodyweightgain and feed conversion ratio of both groups during
the finisher phase were approximately the same,and the cages housing chicks in control and experimental groups during the finishing phase were settled
separately at both ends in an open-type poultry
house in which environmental conditions were
almost uniform.
Abnormalities in health condition for chicks in
the SL and SL-F groups were not observed, and thelivability did not differ significantly between thetwo groups. Moreover, significant differences inserum biochemical and hematological values of chicks
examined on the last day of the experiment were
not found between the SL and SL-F groups, and noabnormalities were observed after histopathologicalobservation or in necropsy findings for chicks in eachgroup.
Although some feeding experiments of GM corn,
excluding SL, have been done with broiler chicks,
there was no difference in broiler performance owing
to the consumption of the GM corn compared withthe conventional non-GM corn (Brake & Vlachos
1998; Sidhu et al. 2000; Einspanier et al. 2001).
In this study, the cry9C gene and Cry9C protein were
not detected by PCR and ELISA in blood, liver andmuscle of chicks at any sampling period. Duggan et al.
(2000) showed that corn chromosomal DNA, including a 1914-bp fragment of the cry1A (b) gene was

Animal Science Journal (2002) 73, 221–228

degraded after just a 1-min incubation with ovine
rumen fluid. Thus, there is a possibility that the Cry9C
protein was degraded in the gastrointestinal tract ofbroiler chicks and was not transferred to blood, liver
and muscles.
In conclusion, the results obtained in this study
indicate that feeding a diet containing SL up to thenutritional upper limit of the formulation, does not
influence growth performance, health condition andphysiological function in broiler chicks between
hatching and the end of fattening, compared withchicks fed on a diet containing SL-F. The cry9C gene
and Cry9C protein are not transferred to blood, liver
and muscles of broiler chicks.

ACKNOWLEDGMENT
This study was funded by a grant from the Agriculture
Livestock Industry Corporation.

REFERENCES
Anon. 1980. A Color Atlas of Hematology of Domestic Animals.
Chikusan Publishing Co, Ltd., Tokyo. (in Japanese).
Brake J, Vlachos D. 1998. Evaluation of transgenic event
176 ‘Bt’ corn in broiler chickens. Poultry Science 77, 648–
653.
Duggan PS, Chambers PA, Heritage J, Forbes JM. 2000.
Survival of free DNA encoding antibiotic resistance fromtransgenic maize and the transformation activity of DNA
in bovine saliva, bovine rumen fluid and silage effluent.
FEMS Microbiology Letters 191, 71–77.

227

C. YONEMOCHI et al.

Einspanier R, Klotz A, Kraft J, Aulrich K, Poser R, Schwagele
F, Jahreis G, Flachowsky G. 2001. The fate of forage plant
DNA in farm animals: a collaborative case-study investigating cattle and chicken fed recombinant plant material.
European Food Research and Technology 212, 129–134.
Ministry of Agriculture, Forestry and Fisheries ResearchCouncil Secretariat, MAFF. 1995. Standard Table of Feed
Composition in Japan (1995). Central Association ofLivestock Industry, Tokyo. (in Japanese).
Ministry of Agriculture, Forestry and Fisheries ResearchCouncil Secretariat, MAFF. 1996. Guidelines for Safety
Assessment of Feed Produced by the Recombitant DNA
Techniques. Central Association of Livestock Industry,
Tokyo. (in Japanese).
Ministry of Agriculture, Forestry and Fisheries ResearchCouncil Secretariat, MAFF. 1997. Japanese FeedingStandard for Poultry (1997). Central Association ofLivestock Industry, Tokyo. (in Japanese).
Organisation for Economic Co-operation and Development

Animal Science Journal (2002) 73, 221–228(OECD). 1993. Safety Evaluation of Foods Derived byModern Biotechnology; Concepts and Principles. In:
OECD Documents. OECD, Paris.
Schepf E, Crickmore N, Van Rie J, Lereclus D, Baum J,
Feitelson J, Zeigler DR, Dean DH. 1998. Bacillus
thuringiensis and its pesticidal crystal proteins. Microbiology
and Molecular Biology Reviews 78, 775–806.
Shen XJ, Suzuki H, Tsudzuki M, Ito S, Nakamura T. 1999.
Comparison of cytochrome b region among Chinese
painted quail and white broiler chicken based on PCRRFLP analysis. Journal of Poultry Science 36, 287–294.
Sidhu R, Hammond BG, Fuchs RL, Mutz JN, Holden LR,
George B, Olson T. 2000. Glyphosate-tolerant corn: The
composition and feeding value of gain from glyphosatetolerant corn is equivalent to that of conventional corn
(Zea maize L.). Journal of Agricultural and Food Chemistry48, 2305–2312.
Yoshida M. 1985. Design of Experiments for Animal Husbandry.
Yokendo Ltd, Tokyo. (in Japanese).

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